KR100484653B1 - Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor - Google Patents

Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor Download PDF

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Publication number
KR100484653B1
KR100484653B1 KR1020040031977A KR20040031977A KR100484653B1 KR 100484653 B1 KR100484653 B1 KR 100484653B1 KR 1020040031977 A KR1020040031977 A KR 1020040031977A KR 20040031977 A KR20040031977 A KR 20040031977A KR 100484653 B1 KR100484653 B1 KR 100484653B1
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KR
South Korea
Prior art keywords
prokaryotes
cistron
sequence encoding
dna sequence
target protein
Prior art date
Application number
KR1020040031977A
Other languages
Korean (ko)
Inventor
이봉용
박승국
박신혜
권오병
윤채하
김영주
김송영
민경현
이윤석
이태희
구태영
Original Assignee
주식회사 대웅
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Publication date
Application filed by 주식회사 대웅 filed Critical 주식회사 대웅
Priority to KR1020040031977A priority Critical patent/KR100484653B1/en
Priority to JP2007511267A priority patent/JP4510884B2/en
Priority to US11/579,753 priority patent/US7811786B1/en
Priority to AT04736706T priority patent/ATE468398T1/en
Priority to DE602004027310T priority patent/DE602004027310D1/en
Priority to EP04736706A priority patent/EP1747274B9/en
Priority to PCT/KR2004/001393 priority patent/WO2005108585A1/en
Priority to CN200480043539XA priority patent/CN101023174B/en
Application granted granted Critical
Publication of KR100484653B1 publication Critical patent/KR100484653B1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli

Abstract

Provided are a method for producing active and soluble proteins in prokaryotes and polycistronic vectors therefor, thereby producing foreign proteins having activity contained in the inclusion body in prokaryotes. As a result, a large quantity of foreign proteins having activity and solubility are produced from prokaryotes. The method for producing active and soluble proteins in prokaryotes comprises the steps of: (1) preparing a recombinant vector comprising (i) a promoter operable in prokaryotes, (ii) a first cistron containing the DNA sequence encoding a target protein, and (iii) a second cistron containing the DNA sequence encoding beta-lactamase, in which the target protein and beta-lactamase are expressed in a type of polycistron in the first cistron and second cistron, respectively, and polycistron is operably linked in a direction from 5' to 3' to the promoter; and (2) transforming prokaryote cells with the recombinant vector to produce transformants; and (3) culturing the transformants of step (2), and recovering the target protein from the cultured medium, wherein the prokaryote is Escherichia coli. The polycistronic vector comprises (i) a promoter operable in prokaryotes, (ii) a first cistron containing the DNA sequence encoding a target protein, and (iii) a second cistron containing the DNA sequence encoding beta-lactamase, which are operably linked from 5' to 3'.
KR1020040031977A 2004-05-06 2004-05-06 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor KR100484653B1 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
KR1020040031977A KR100484653B1 (en) 2004-05-06 2004-05-06 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor
JP2007511267A JP4510884B2 (en) 2004-05-06 2004-06-11 Method for producing active soluble protein in prokaryotic cells and polycistron vector therefor
US11/579,753 US7811786B1 (en) 2004-05-06 2004-06-11 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor
AT04736706T ATE468398T1 (en) 2004-05-06 2004-06-11 MANUFACTURING PROCESS FOR THE PRODUCTION OF ACTIVE AND SOLUBLE PROTEINS IN PROKARYONTS AND POLYCISTRONIC VECTORS THEREFOR
DE602004027310T DE602004027310D1 (en) 2004-05-06 2004-06-11 METHOD OF MANUFACTURING ACTIVE AND SOLUBLE PROTEINS IN PROKARYONS AND POLYCISTRONIC VECTORS THEREFOR
EP04736706A EP1747274B9 (en) 2004-05-06 2004-06-11 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor
PCT/KR2004/001393 WO2005108585A1 (en) 2004-05-06 2004-06-11 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor
CN200480043539XA CN101023174B (en) 2004-05-06 2004-06-11 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1020040031977A KR100484653B1 (en) 2004-05-06 2004-05-06 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor

Publications (1)

Publication Number Publication Date
KR100484653B1 true KR100484653B1 (en) 2005-04-20

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Application Number Title Priority Date Filing Date
KR1020040031977A KR100484653B1 (en) 2004-05-06 2004-05-06 Preparation method for the production of active and soluble proteins in prokaryotes and polycistronic vectors therefor

Country Status (8)

Country Link
US (1) US7811786B1 (en)
EP (1) EP1747274B9 (en)
JP (1) JP4510884B2 (en)
KR (1) KR100484653B1 (en)
CN (1) CN101023174B (en)
AT (1) ATE468398T1 (en)
DE (1) DE602004027310D1 (en)
WO (1) WO2005108585A1 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
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KR101477235B1 (en) 2012-09-28 2015-01-02 선문대학교 산학협력단 Construction of a Multi-monocistronic Vector Applicable for Synthetic Biology and Use Thereof
KR20220028520A (en) * 2020-08-28 2022-03-08 한국해양과학기술원 Thermally stable fgf7 polypeptide and use of the same

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CN101743306A (en) 2007-03-23 2010-06-16 威斯康星校友研究基金会 Somatic cell reprogramming
ES2587395T3 (en) 2008-06-04 2016-10-24 Cellular Dynamics International, Inc. Procedures for the production of IPS cells using a non-viral approach
WO2010019569A1 (en) 2008-08-12 2010-02-18 Cellular Dynamics International. Inc. Methods for the production of ips cells
JP2013511261A (en) * 2009-11-18 2013-04-04 マリー ゴールバーン シーオー−オペレイティブ シーオー．リミテッド Recombinant microorganism
GB201018664D0 (en) * 2010-11-05 2010-12-22 Msd Biolog Uk Ltd Expression process
CN102329811B (en) * 2011-09-15 2014-08-13 生工生物工程（上海）股份有限公司 Ampicillin resistance plasmid vector as well as preparation and application thereof
HUP1200171A1 (en) * 2012-03-19 2013-09-30 Richter Gedeon Nyrt Methods for the production of polypeptides
WO2014082080A2 (en) * 2012-11-26 2014-05-30 Callidus Biopharma, Inc. Methods for coupling targeting peptides onto recombinant lysosomal enzymes for improved treatments of lysosomal storage diseases
AU2015308897B2 (en) * 2014-08-28 2021-03-04 Theriva Biologics, Inc. E. coli-based production of beta-lactamase
CN104946647A (en) * 2015-06-08 2015-09-30 南京理工大学 Chlamydomonas reinhardtii intergenic expression treating element
CN105420263B (en) * 2015-12-16 2019-07-26 温州医科大学 The production method of recombinant human fibroblast growth factor -17
CN106434733B (en) * 2016-08-01 2019-09-27 江南大学 A kind of expression vector and its application suitable for Corynebacterium glutamicum
JP7083124B2 (en) 2017-04-25 2022-06-10 国立大学法人弘前大学 Muconic acid-producing transformed microorganisms and their utilization
SG11201912790WA (en) * 2017-06-23 2020-01-30 Zhuhai Essex Bio Pharmaceutical Co Ltd Method for producing soluble recombinant human-basic fibroblast growth factor (rh-bfgf)
KR102480402B1 (en) * 2017-06-23 2022-12-22 주하이 에섹스 바이오-파머슈티컬 코., 엘티디. A pharmaceutical composition comprising recombinant human-basic fibroblast growth factor (rh-bFGF) and rh-bFGF

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IL80511A0 (en) 1985-11-05 1987-02-27 Genetics Inst Multiply amplifiable vectors for high level expression of exogenous dna
JP2570651B2 (en) * 1986-10-09 1997-01-08 藤沢薬品工業株式会社 Method for producing Met-insulin-like growth factor I
US5648254A (en) 1988-01-15 1997-07-15 Zymogenetics, Inc. Co-expression in eukaryotic cells
US6027888A (en) 1996-04-05 2000-02-22 Board Of Regents, The University Of Texas System Methods for producing soluble, biologically-active disulfide-bond containing eukaryotic proteins in bacterial cells
KR100203919B1 (en) 1996-10-04 1999-06-15 신동권 A novel expression vector which produces water soluble protein
KR100254063B1 (en) 1997-09-30 2000-04-15 신동권 Process for preparing recombinant proteins using recombinant e. coli
AU2002217505B2 (en) 2000-12-26 2006-03-02 Sekisui Chemical Co., Ltd. Process for producing recombinant protein and fused protein
IL157378A0 (en) * 2001-03-09 2004-02-19 Genentech Inc Process for production of polypeptides
KR20040007892A (en) 2002-07-11 2004-01-28 주식회사 바이오프로젠 Recombinant Human Ferritin Protein And A Process for Producing the Same

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101477235B1 (en) 2012-09-28 2015-01-02 선문대학교 산학협력단 Construction of a Multi-monocistronic Vector Applicable for Synthetic Biology and Use Thereof
KR20220028520A (en) * 2020-08-28 2022-03-08 한국해양과학기술원 Thermally stable fgf7 polypeptide and use of the same
KR102440312B1 (en) 2020-08-28 2022-09-05 한국해양과학기술원 Thermally stable fgf7 polypeptide and use of the same

Also Published As

Publication number Publication date
EP1747274B1 (en) 2010-05-19
EP1747274A4 (en) 2007-07-11
CN101023174B (en) 2011-03-02
US7811786B1 (en) 2010-10-12
WO2005108585A1 (en) 2005-11-17
EP1747274B9 (en) 2011-02-02
JP2007535923A (en) 2007-12-13
JP4510884B2 (en) 2010-07-28
CN101023174A (en) 2007-08-22
ATE468398T1 (en) 2010-06-15
DE602004027310D1 (en) 2010-07-01
EP1747274A1 (en) 2007-01-31

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